Division of Gastroenterology, Department of Medicine Department of Digestive Disease Information & Research, Kurume University School of Medicine, Kurume, Japan.
Hepatol Res. 2011 Nov;41(11):1120-5. doi: 10.1111/j.1872-034X.2011.00863.x.
Serum albumin exists in both oxidized and reduced forms. Reduced albumin shows higher antioxidative effects; however, the percentage of reduced albumin is low in human serum albumin (HSA) preparation. Stronger neo-minophagen C (SNMC) containing cysteine, a nucleophilic amino acid, has the potential to control the redox state of albumin. The aim of this study was to develop a method for increasing the fraction of reduced albumin in HAS preparation using SNMC.
Human serum albumin preparations were purchased from four different manufacturers. As a reducing agent, SNMC (50, 100, or 200 µL) was added to 62.5 mg of each HSA preparation, and the mixture was incubated at room temperature for 10-480 min. The percentages of reduced albumin were determined by high-performance liquid chromatography.
The percentage of reduced albumin in the HSA preparation significantly increased 15 min after treatment with 200 µL of SNMC (manufacturer A; 27.7 ± 0.18% vs. 78.7 ± 0.36%, P < 0.01), and a dose-dependent relationship was observed between the increase in the percentage of reduced albumin and dose of SNMC. Similar results were obtained with the other three HSA preparations. The percentage of reduced albumin reached its peak at 15 min after mixing SNMC with an HSA preparation, and then gradually decreased with duration, irrespective of the dose of SNMC.
We devised a method for increasing the reduced albumin fraction in an HSA preparation by using SNMC. We also determined the time-and dose-differences in the effect of SNMC on redox state of HSA.
血清白蛋白存在氧化型和还原型两种形式。还原型白蛋白具有更高的抗氧化作用；然而，人血清白蛋白（HSA）制剂中的还原型白蛋白比例较低。含有亲核氨基酸半胱氨酸的更强的新清蛋白 C（SNMC）有可能控制白蛋白的氧化还原状态。本研究旨在开发一种使用 SNMC 增加 HSA 制剂中还原型白蛋白比例的方法。
从四个不同的制造商购买人血清白蛋白制剂。将 SNMC（50、100 或 200μL）作为还原剂添加到 62.5mg 的每种 HSA 制剂中，并在室温下孵育 10-480 分钟。通过高效液相色谱法测定还原型白蛋白的百分比。
用 200μL SNMC 处理 15 分钟后，HSA 制剂中的还原型白蛋白百分比显著增加（制造商 A；27.7±0.18%对 78.7±0.36%，P<0.01），并且观察到还原型白蛋白百分比的增加与 SNMC 剂量之间存在剂量依赖性关系。其他三种 HSA 制剂也得到了类似的结果。SNMC 与 HSA 制剂混合 15 分钟后，还原型白蛋白的百分比达到峰值，然后随着时间的推移逐渐下降，与 SNMC 的剂量无关。
我们设计了一种使用 SNMC 增加 HSA 制剂中还原型白蛋白比例的方法。我们还确定了 SNMC 对 HSA 氧化还原状态影响的时间和剂量差异。
